Lipidomics in melanoma: identification of new lipid prognostic biomarkers

Los capítulos de Resultados, Discusion y Conclusiones están sujetos a confidencialidad. 179 p.El objetivo de esta tesis doctoral fue el de encontrar nuevos biomarcadores para el melanoma, ya que lasupervivencia de los pacientes se reduce dramáticamente cuando no se detecta en la fase temprana de laenfermedad. Para esto, se ha estudiado y comparado el contenido lipídico de numerosas líneas celularesde melanocitos de piel sanos, melanocitos extraidos de nevus, melanoma primarios y melanomametastásico, aplicando diferentes metodologías de análisis lipidómico. Los resultados demuestran que lascélulas sanas y tumorales presenta una huella lipídica diferencial. Además, aplicando diferentes análisisestadísticos, hemos detectado numerosas especies de lípidos que presentan una alteración en suintensidad al comparar los diferentes grupos de estudio. Todas estas especies lipídicas se han definidocomo potenciales biomarcadores, y la metodología de detectarlos va a ser próximamente patentada. Porotro lado, hemos comprobado que algunas de las enzimas que catabolizan algunos de los lípidos conexpresión diferencial en melanoma tienen su expresión y actividad alteradas. En concreto, hemoscomprobado que la fosfolipasa D2 ayuda en el proceso tumoral y metastásico del melanoma

High resolution and fidelity 3D printing of Laponite and alginate ink hydrogels for tunable biomedical applications

The formulation of hydrogels that meet the necessary flow characteristics for their extrusion-based 3D printing while providing good printability, resolution, accuracy and stability, requires long development processes. This work presents the technological development of a hydrogel-based ink of Laponite and alginate and evaluates its printing capacity. As a novelty, this article reports a standardizable protocol to quantitatively define the best printing parameters for the development of novel inks, providing new printability evaluation parameters such as the Printing Accuracy Escalation Index. As a result, this research develops a printable Laponite-Alginate hydrogel that presents printability characteristics. This ink is employed for the reproducible manufacture of 3D printed scaffolds with versatile and complex straight or curved printing patterns for a better adaptation to different final applications. Obtained constructs prove to be stable over time thanks to the optimization of a curing process. In addition, the study of the swelling and degradation behavior of the Laponite and alginate 3D printed scaffolds in different culture media allows the prediction of their behavior in future in vitro or in vivo developments. Finally, this study demonstrates the absence of cytotoxicity of the printed formulations, hence, setting the stage for their use in the field of biomedicine.This project has been partially supported by Spanish government Ministerio de Ciencia e Innovación Grant PID2021-122577OB-I00 funded by MCIN/AEI/ 10.13039/501100011033 and by “ERDF A way of making Europe”. Grant IT1448-22 funded by Basque Government and Fundación Vital Fundazioa (vital21/28). Elena Munoz-Perez thanks the Basque Government for the predoctoral grant (PRE_2022_2_0115). Arantza Perez-Valle thanks the Spanish Government for the postdoctoral grant (MARSA55/21)

Upregulated phospholipase D2 expression and activity is related to the metastatic properties of melanoma

[EN] The incidence rates of melanoma have increased steadily in recent decades and nearly 25% of the patients diagnosed with early-stage melanoma will eventually develop metastasis, for which there is currently no fully effective treatment. The link between phospholipases and tumors has been studied extensively, particularly in breast and colon cancers. With the aim of finding new biomarkers and therapeutic options for melanoma, the expression of different phospholipases was assessed in 17 distinct cell lines in the present study, demonstrating that phospholipase D2 (PLD2) is upregulated in metastatic melanoma as compared to normal skin melanocytes. These results were corroborated by immunofluorescence and lipase activity assays. Upregulation of PLD2 expression and increased lipase activity were observed in metastatic melanoma relative to normal skin melanocytes. So far, the implication of PLD2 activity in melanoma malignancies has remained elusive. To the best of our knowledge, the present study was the first to demonstrate that the overexpression of PLD2 enhances lipase activity, and its effect to increase the proliferation, migration and invasion capacity of melanoma cells was assessed with XTT and Transwell assays. In addition, silencing of PLD2 in melanoma cells reduced the metastatic potential of these cells. The present study provided evidence that PLD2 is involved in melanoma malignancy and in particular, in its metastatic potential, and established a basis for future studies evaluating PLD2 blockade as a therapeutic strategy to manage this condition.This study was supported by grants from the University of the Basque Country/EHU (grant no. GIU17/066) and Ministerio de Economia y Competividad MINECO-ONCOFINDER of the Spanish Government (grant no. RTC.2015-3693-1)

Uso de localizadores para personas mayores

3rd International Conference on the Elderly and New Technologies. III Jornadas Internacionales de Mayores y Nuevas Tecnologías.Descriptive cross-sectional study which uses self-administered surveys to evaluate the data from three independent samples: professionals, relatives of users of the tracking device and relatives of users who do no use it. Methods: specific survey for 30 professionals. For 7 relatives of users who make use of the device: Quebec survey (QUEST 2.0), Zarit scale and caregiver strain index. For 7 relatives of users who do not make use of the device: Zarit Scale and caregiver strain index. Results: 20% of the professionals are aware of the existence of some kind of gps device, only 13.3% can specify the sort of gps. The benefits with the highest score in the Liker scale from 0 to 5 are the tranquillity of the family (4.60), followed by the need of information (4.46), and the item with the lowest score is the tranquillity of the user (3.79). Among those patients carrying a tracking device, 85.71% are men, whereas this percentage turns to a 71,43% among those who do not carry a tracking device. The burden degree of caregivers is an average of 77.86% in those who do not use a tracking device, whereas it goes down to 66.58% in those who do. Caregivers using the tracking device get a 57.1% of greater burden, and a 14.3% of minimum burden. Caregivers who do not use the tracking device get a 100% of greater burden. In the strain index test the results show that a 92.9% of caregivers reach high levels of burden, whereas for 7.1% there was no strain at all. 84.6% of caregivers are women. Conclusions: professionals do not know the tracking device, and the burden of caregivers decreases when they use it. The use of the tracking device should be discussed in an ethical debate by professionals of all areas and relatives of patients.Estudio descriptivo transversal que evalúa, mediante encuestas autoadministrada, los datos procedentes de tres muestras independientes: profesionales, familiares de usuarios que utilizan el dispositivo de localización y familiares de usuarios que no lo utilizan. Métodos: encuesta especifica para 30 profesionales. Para 7 familiares de usuarios que utilizan el dispositivo: encuesta Quebec (QUEST 2.0), escala Zarit y el índice de esfuerzo del cuidador. Para 7 familiares que no utilizan el dispositivo: la escala Zarit y el Índice de esfuerzo del cuidador. Resultados: el 20 % de los profesionales admite conocer algún dispositivo gps, tan solo un 13,3 % saben especificar el tipo de gps. Los beneficios mejor puntuados en la escala Liker 0 a 5 son tranquilidad de la familia (4,60), seguido de la necesidad de información (4,46), y el ítem menos valorado la tranquilidad del usuario (3,79). Los pacientes que llevan localizador, los hombres representan un 85,71 % y en los que no llevan localizador representan un 71,43 %. El grado de sobrecarga que tienen los cuidadores obtiene una media de 77,86 % para los que no utilizan el localizador, frente a un 66,58 % para los que sí utilizan el localizador. Los cuidadores que utilizan el dispositivo obtienen un 57,1 % de sobrecarga intensa, un 14,3% sobrecarga leve. Los cuidadores que no utilizan el localizador obtienen un 100 % de sobrecarga intensa. En los resultados del test de índice de esfuerzo, el 92,9 % de los cuidadores tienen puntuaciones elevadas de esfuerzo, frente al 7,1 % que no presentan esfuerzo. De los cuidadores, el 84,6 % son mujeres. Conclusiones: los profesionales no conocen el dispositivo y la sobrecarga en los cuidadores disminuye con el uso del localizador. Debería plantearse un debate ético entre los diferentes colectivos de profesionales y familiares sobre su uso

Plasma-Based Bioinks for Extrusion Bioprinting of Advanced Dressings

Extrusion bioprinting based on the development of novel bioinks offers the possibility of manufacturing clinically useful tools for wound management. In this study, we show the rheological properties and printability outcomes of two advanced dressings based on platelet-rich plasma (PRP) and platelet-poor plasma (PPP) blended with alginate and loaded with dermal fibroblasts. Measurements taken at 1 h, 4 days, and 18 days showed that both the PRP- and PPP-based dressings retain plasma and platelet proteins, which led to the upregulation of angiogenic and immunomodulatory proteins by embedded fibroblasts (e.g., an up to 69-fold increase in vascular endothelial growth factor (VEGF), an up to 188-fold increase in monocyte chemotactic protein 1 (MCP-1), and an up to 456-fold increase in hepatocyte growth factor (HGF) 18 days after printing). Conditioned media harvested from both PRP and PPP constructs stimulated the proliferation of human umbilical vein endothelial cells (HUVECs), whereas only those from PRP dressings stimulated HUVEC migration, which correlated with the VEGF/MCP-1 and VEGF/HGF ratios. Similarly, the advanced dressings increased the level of interleukin-8 and led to a four-fold change in the level of extracellular matrix protein 1. These findings suggest that careful selection of plasma formulations to fabricate wound dressings can enable regulation of the molecular composition of the microenvironment, as well as paracrine interactions, thereby improving the clinical potential of dressings and providing the possibility to tailor each composition to specific wound types and healing stages.This work was fully supported by a collaborative fundamental research grant from the Basque Government Elkartek program under grant nº. B4H KK-2019-0006-BC

The Multifunctional Role of SPANX-A/D Protein Subfamily in the Promotion of Pro-Tumoural Processes in Human Melanoma

Human sperm protein associated with the nucleus on the X chromosome (SPANX) genes encode a protein family (SPANX-A, -B, -C and -D), whose expression is limited to the testis and spermatozoa in normal tissues and various tumour cells. SPANX-A/D proteins have been detected in metastatic melanoma cells, but their contribution to cancer development and the underlying molecular mechanisms of skin tumourigenesis remain unknown. Combining functional and proteomic approaches, the present work describes the presence of SPANX-A/D in primary and metastatic human melanoma cells and how it promotes pro-tumoural processes such as cell proliferation, motility and migration. We provide insights into the molecular features of skin tumourigenesis, describing for the first time a multifunctional role of the SPANX-A/D protein family in nuclear function, energy metabolism and cell survival, considered key hallmarks of cancer. A better comprehension of the SPANX-A/D protein subfamily and its molecular mechanisms will help to describe new aspects of tumour cell biology and develop new therapeutic targets and tumour-directed pharmacological drugs for skin tumoursUniversity of the Basque Country (UPV/EHU) (GIU19/018). IU-A is supported by a fellowship from the University of the Basque Country (UPV/EHU). IM-H is supported by a fellowship from the Basque Government

A UHPLC-Mass Spectrometry View of Human Melanocytic Cells Uncovers Potential Lipid Biomarkers of Melanoma

Melanoma is the deadliest form of skin cancer due to its ability to colonize distant sites and initiate metastasis. Although these processes largely depend on the lipid-based cell membrane scaffold, our understanding of the melanoma lipid phenotype lags behind most other aspects of this tumor cell. Here, we examined a panel of normal human epidermal and nevus melanocytes and primary and metastatic melanoma cell lines to determine whether distinctive cell-intrinsic lipidomes can discern non-neoplastic from neoplastic melanocytes and define their metastatic potential. Lipidome profiles were obtained by UHPLC-ESI mass-spectrometry, and differences in the signatures were analyzed by multivariate statistical analyses. Significant and highly specific changes in more than 30 lipid species were annotated in the initiation of melanoma, whereas less numerous changes were associated with melanoma progression and the non-malignant transformation of nevus melanocytes. Notably, the “malignancy lipid signature” features marked drops in pivotal membrane lipids, like sphingomyelins, and aberrant elevation of ether-type lipids and phosphatidylglycerol and phosphatidylinositol variants, suggesting a previously undefined remodeling of sphingolipid and glycerophospholipid metabolism. Besides broadening the molecular definition of this neoplasm, the different lipid profiles identified may help improve the clinical diagnosis/prognosis and facilitate therapeutic interventions for cutaneous melanoma.This research was funded in part by grants from the Ministry of Economy; Industry and Competitiveness (RTC-2015-3693-1); Ministry of Science and Innovation (RTI-2018-095134-B-I00); Basque Government (IT971-16; IT1162-19; KK2016-036; KK2017-041 and KK2018-00090) and UPV/EHU (GIU17/066)

Treatment variability and its relationships to outcomes among patients with Wernicke's encephalopathy: A multicenter retrospective study

Background: Despite guidelines and recommendations, Wernicke's encephalopathy (WE) treatment lacks evidence, leading to clinical practice variability.Aims: Given the overall lack of information on thiamine use for WE treatment, we analyzed data from a large, well-characterized multicenter sample of patients with WE, examining thiamine dosages; factors associated with the use of different doses, frequencies, and routes; and the influence of differences in thiamine treatment on the outcome.Methods: This retrospective study was conducted with data from 443 patients from 21 centers obtained from a nationwide registry of the Spanish Society of Internal Medicine (from 2000 to 2012). Discharge codes and Caine criteria were applied for WE diagnosis, and treatment-related (thiamine dosage, frequency, and route of administration) demographic, clinical, and outcome variables were analyzed.Results: We found marked variability in WE treatment and a low rate of high-dose intravenous thiamine administration. Seventy-eight patients out of 373 (20.9%) received > 300 mg/day of thiamine as initial dose. Patients fulfilling the Caine criteria or presenting with the classic WE triad more frequently received parenteral treatment. Delayed diagnosis (after 24 h hospitalization), the fulfillment of more than two Caine criteria at diagnosis, mental status alterations, and folic acid deficiency were associated significantly with the lack of complete recovery. Malnutrition, reduced consciousness, folic acid deficiency, and the lack of timely thiamine treatment were risk factors for mortality.Conclusions: Our results clearly show extreme variability in thiamine dosages and routes used in the management of WE. Measures should be implemented to ensure adherence to current guidelines and to correct potential nutritional deficits in patients with alcohol use disorders or other risk factors for WE

Juan Labranz cave: a Quaternary deposit in the central Iberian Peninsula

Here we report the preliminary results from the 2015–19 s prospecting explorations, excavations, and research from Juan Labranz Cave, a new Quaternary palaeontological deposit rich in mammals that offers information on the faunal context of the southern sub-plateau. This cave is located on the border of the Iberian range, at the Sierra de Valdecabras, Cuenca, at 1.279 metres above sea level. This study includes the first georeferenced digital map of the cave and a preliminary analysis of its chronology, palynology, macro- and microvertebrate palaeontology, and taphonomy. The cave is interpreted as a hyena den, and this would represent one of the highest elevation cavities where the activity of this taxon is recorded. Moreover, we consider this site important and unique because it constitutes one of the very few Pleistocene cave sites in the southern sub-plateau. It is strategically located on the border between the Iberian range and the Tajo Tertiary Basin, at the Júcar River valley, which represents the only great natural corridor that covers hundreds of kilometres and connects two very important palaeoecological areas, the interior of the Iberian Peninsula and the Mediterranean basin

Treatment variability and its relationships to outcomes among patients with Wernicke's encephalopathy : A multicenter retrospective study

CatedresBackground: Despite guidelines and recommendations, Wernicke's encephalopathy (WE) treatment lacks evidence, leading to clinical practice variability. Aims: Given the overall lack of information on thiamine use for WE treatment, we analyzed data from a large, well-characterized multicenter sample of patients with WE, examining thiamine dosages; factors associated with the use of different doses, frequencies, and routes; and the influence of differences in thiamine treatment on the outcome. Methods: This retrospective study was conducted with data from 443 patients from 21 centers obtained from a nationwide registry of the Spanish Society of Internal Medicine (from 2000 to 2012). Discharge codes and Caine criteria were applied for WE diagnosis, and treatment-related (thiamine dosage, frequency, and route of administration) demographic, clinical, and outcome variables were analyzed. Results: We found marked variability in WE treatment and a low rate of high-dose intravenous thiamine administration. Seventy-eight patients out of 373 (20.9%) received > 300 mg/day of thiamine as initial dose. Patients fulfilling the Caine criteria or presenting with the classic WE triad more frequently received parenteral treatment. Delayed diagnosis (after 24 h hospitalization), the fulfillment of more than two Caine criteria at diagnosis, mental status alterations, and folic acid deficiency were associated significantly with the lack of complete recovery. Malnutrition, reduced consciousness, folic acid deficiency, and the lack of timely thiamine treatment were risk factors for mortality. Conclusions: Our results clearly show extreme variability in thiamine dosages and routes used in the management of WE. Measures should be implemented to ensure adherence to current guidelines and to correct potential nutritional deficits in patients with alcohol use disorders or other risk factors for WE